Preventive Effect of Vitamin C on Dextran Sulfate Sodium (DSS)-Induced Colitis via the Regulation of IL-22 and IL-6 Production in Gulo(-/-) Mice.

Reactive oxygen species (ROS), which are exceptionally high in IBD lesions, are known to cause abnormal immune responses to inflammatory reactions in inflammatory bowel diseases (IBD) through damage to the intestinal mucosal linings. Moreover, they are theorized to be an agent of IBD development. Vitamin C is widely known to be an effective antioxidant for its ability to regulate inflammatory responses through its ROS scavenging effect. Therefore, we examined vitamin C's influence on the development and progression of IBD in Gulo(-/-) mice, which cannot synthesize vitamin C like humans due to a defect in the expression of L-gulono-γ-lactone oxidase, an essential enzyme for vitamin C production. First, we found extensive oxidative stress and an inflammation increase in the colon of vitamin C-insufficient Gulo(-/-) mice. We also found decreased IL-22 production and NKp46(+) cell recruitment and the impaired activation of the p38MAPK pathway. Additionally, comparing vitamin C-insufficient Gulo(-/-) mice to vitamin C-sufficient Gulo(-/-) mice and wild-type mice, the insufficient group faced a decrease in mucin-1 expression, accompanied by an increase in IL-6 production, followed by the activation of the STAT3 and Akt pathways. The results suggest that vitamin C insufficiency induces severe colitis, meaning vitamin C could also take on a preventative role by regulating the production of cytokines and the induction of inflammation.

Comparative transcriptome analysis revealed omnivorous adaptation of the small intestine of Melinae.

As the main digestive organ, the small intestine plays a vital role in the digestion of animals. At present, most of the research on animal feeding habits focuses on carnivores and herbivores. However, the mechanism of feeding and digestion in omnivores remains unclear. This study aims to reveal the molecular basis of the omnivorous adaptive evolution of Melinae by comparing the transcriptome of the small intestines of Asian Badgers (Meles leucurus) and Northern Hog Badgers (Arctonyx albogularis). We obtained high-quality small intestinal transcriptome data from these two species. Key genes and signalling pathways were analysed through Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and other databases. Research has mainly found that orthologous genes related to six enzymes have undergone adaptive evolution. In addition, the study also found three digestion-related pathways (cGMP-PKG, cAMP, and Hippo). They are related to the digestion and absorption of nutrients, the secretion of intestinal fluids, and the transport of food through the small intestine, which may help omnivorous animals adapt to an omnivorous diet. Our study provides insight into the adaptation of Melinae to omnivores and affords a valuable transcriptome resource for future research.

BIOACCUMULATION OF MERCURY IN A TERRESTRIAL CARNIVORE, AMERICAN MARTEN (MARTES AMERICANA).

Studies that assess mercury bioaccumulation in small carnivores in terrestrial habitats are limited. We quantified total mercury (THg) in American marten (Martes americana) that were harvested for fur in Michigan, US, during 2013 and 2014. We quantified THg (dry weight) in hair, kidney, and liver samples and further analyzed hair for potential demographic and ecological factors that influence THg bioaccumulation. We found THg concentrations to be the highest in hair (1.228±0.475 µg/g, n=40), followed by kidney (0.922±0.651 µg/g, n=29) and liver (0.344±0.219 µg/g, n=26). Total mercury distributed predictably and significantly between tissue types, and hair was moderately predictive at modeling THg in kidney (R2=0.50, P<0.001, n=29) and weakly predictive in liver (R2=0.35, P<0.001, n=26), suggesting that hair, which is easily obtained, could be a useful sample type for future biomonitoring programs. The concentrations of THg in hair were higher in adults relative to juveniles, and adult female martens had the highest levels of THg (1.980±0.188 µg/g), as compared to juveniles and adult males. Results of generalized linear modeling suggested that THg hair concentrations were positively associated with marten age and trophic position (stable isotope ratio, δ15N). An interaction between δ15N and the year marten carcasses were collected showed that δ15N alone could be highly predictive of THg in some years but not in others. Annual changes in diet could lead to differing rates of mercury bioaccumulation and alter the usefulness of δ15N to predict THg in marten tissues. Further research should explore the connections between changes in prey availability, types of prey consumed, and the influence on bioaccumulation rates of mercury in terrestrial system mesocarnivores.

Adaptive evolution of the ACSL gene family in Carnivora.

Carnivores exhibit various fat contents and energy reserves to adapt to their environments. However, the molecular mechanisms underlying lipid metabolic differences among carnivores have not been well explored. Long-chain acyl-CoA synthetases (ACSLs) catalyze the initial step in lipid metabolism by activating fatty acids (FAs), and they drive acyl-CoAs toward anabolic lipid synthesis or catabolic ß-oxidation. We identified the sequences of the genes of the ACSL family (ACSL1, ACSL3, ACSL4, ACSL5 and ACSL6) in the sable (Martes zibellina) via transcriptome sequencing. The ACSL gene sequences of 13 other carnivores were obtained from NCBI. Phylogenetic results showed that unlike the widely accepted carnivore phylogeny, Canidae and Felidae tend to group together based on ACSL4 and ACSL6. The evolutionary analyses identified a series of positively selected amino acid residues in ACSL1, ACSL4 and ACSL5. Two radical amino acid substitutions detected in sable suggested potential insights into the molecular mechanism underlying the relatively low fat content in this animal. This is the first study to investigate the molecular mechanisms underlying the adaptive evolution of fat metabolism in carnivores. Overall, the ACSL genes were under different evolutionary forces in carnivores, and some genes have undergone adaptive evolution in lipid metabolism.

Badger macrophages fail to produce nitric oxide, a key anti-mycobacterial effector molecule.

The European badger is recognised as a wildlife reservoir for bovine tuberculosis (bTB); the control of which is complex, costly and controversial. Despite the importance of badgers in bTB and the well-documented role for macrophages as anti-mycobacterial effector cells, badger macrophage (bdMφ) responses remain uncharacterised. Here, we demonstrate that bdMφ fail to produce nitric oxide (NO) or upregulate inducible nitric oxide synthase (iNOS) mRNA following Toll-like receptor (TLR) agonist treatment. BdMφ also failed to make NO after stimulation with recombinant badger interferon gamma (bdIFNγ) or a combination of bdIFNγ and lipopolysaccharide. Exposure of bdMφ to TLR agonists and/or bdIFNγ resulted in upregulated cytokine (IL1ß, IL6, IL12 and TNFα) mRNA levels indicating that these critical pathways were otherwise intact. Although stimulation with most TLR agonists resulted in strong cytokine mRNA responses, weaker responses were evident after exposure to TLR9 agonists, potentially due to very low expression of TLR9 in bdMφ. Both NO and TLR9 are important elements of innate immunity to mycobacteria, and these features of bdMφ biology would impair their capacity to resist bTB infection. These findings have significant implications for the development of bTB management strategies, and support the use of vaccination to reduce bTB infection in badgers.

Protection of horse ears against Simulid parasitism: Efficacy of a mammal semiochemical solution over 10hours.

Hematophagous insects can be vectors of pathogens and cause significant economic loss in zootechnical production. Among biting insects, many dipteran species feed on horse blood. The black fly (Diptera: Simuliidae) group, is responsible for several disorders in horses and inflicts painful bites that lead to undesirable behaviours in horses, particularly when bites occur in sensitive areas such as the inner ear. A field study was conducted in a French equestrian center during which a semiochemical was applied on horses' ears to assess repellent efficacy against simulid infestation. During the first phase of the study, efficacy was evaluated over a one hour period. Then, during the second phase of the study, persistency of the effect was tested at 8, 9 and 10h after application. The results of the study's first phase showed 90% efficacy over one hour, with 121.5 insects found in control ears and 12 insects in treated ears (p=0.001). In the second phase of the study, a total amount of 411 insects were observed on control ears whereas only 2 insects were observed on treated ears (p<0.0001); the treatment remained over 98% effective up to 10hours after application. When using a slow release excipient, this semiochemical may offer at least 10h of protection against simulids. This safe, efficient, and long lasting protection could help horses and their owners to manage simulid parasitism.

Acceleration Data Reveal Highly Individually Structured Energetic Landscapes in Free-Ranging Fishers (Pekania pennanti).

Investigating animal energy expenditure across space and time may provide more detailed insight into how animals interact with their environment. This insight should improve our understanding of how changes in the environment affect animal energy budgets and is particularly relevant for animals living near or within human altered environments where habitat change can occur rapidly. We modeled fisher (Pekania pennanti) energy expenditure within their home ranges and investigated the potential environmental and spatial drivers of the predicted spatial patterns. As a proxy for energy expenditure we used overall dynamic body acceleration (ODBA) that we quantified from tri-axial accelerometer data during the active phases of 12 individuals. We used a generalized additive model (GAM) to investigate the spatial distribution of ODBA by associating the acceleration data to the animals' GPS-recorded locations. We related the spatial patterns of ODBA to the utilization distributions and habitat suitability estimates across individuals. The ODBA of fishers appears highly structured in space and was related to individual utilization distribution and habitat suitability estimates. However, we were not able to predict ODBA using the environmental data we selected. Our results suggest an unexpected complexity in the space use of animals that was only captured partially by re-location data-based concepts of home range and habitat suitability. We suggest future studies recognize the limits of ODBA that arise from the fact that acceleration is often collected at much finer spatio-temporal scales than the environmental data and that ODBA lacks a behavioral correspondence. Overcoming these limits would improve the interpretation of energy expenditure in relation to the environment.

Polonium-210 and Caesium-137 in lynx (Lynx lynx), wolverine (Gulo gulo) and wolves (Canis lupus).

Wolves, lynx and wolverines are on the top of the food-chain in northern Scandinavia and Finland. (210)Po and (137)Cs have been analysed in samples of liver, kidney and muscle from 28 wolves from Sweden. In addition blood samples were taken from 27 wolves. In 9 of the wolves, samples of muscle, liver and blood were analysed for (210)Po. Samples of liver and muscle were collected from 16 lynx and 16 wolverines from Norway. The liver samples were analysed for (210)Po and (137)Cs. Only (137)Cs analyses were carried out for the muscle samples. The wolves were collected during the winter 2010 and 2011, while the samples for lynx and wolverines were all from 2011. The activity concentrations of (210)Po in wolves were higher for liver (range 20-523 Bq kg(-1) d.w.) and kidney (range 24-942 Bq kg(-1) d.w.) than muscle (range 1-43 Bq kg(-1) d.w.) and blood (range 2-54 Bq kg(-1) d.w.). Activity ratios, (210)Po/(210)Pb, in wolf samples of muscle, liver and blood were in the ranges 2-77, 9-56 and 2-54. Using a wet weight ratio of 3.8 the maximal absorbed dose from (210)Po to wolf liver was estimated to 3500 µGy per year. Compared to wolf, the ranges of (210)Po in liver samples were lower in lynx (range 22-211 Bq kg(-1) d.w.) and wolverine (range16-160 Bq kg(-1) d.w.). Concentration of (137)Cs in wolf samples of muscle, liver, kidney and blood were in the ranges 70-8410 Bq kg(-1) d.w., 36-4050 Bq kg(-1) d.w., 31-3453 Bq kg(-1) d.w. and 4-959 Bq kg(-1) d.w., respectively. (137)Cs in lynx muscle and liver samples were in the ranges 44-13393 Bq kg(-1) d.w. and 125-10260 Bq kg(-1) d.w. The corresponding values for (137)Cs in wolverine were 22-3405 Bq kg(-1) d.w. for liver and 53-4780 Bq kg(-1) d.w. for muscle. The maximal absorbed dose from (137)Cs to lynx was estimated to 3000 µGy per year.

Mercury in various tissues of three mustelid species and other trace metals in liver of European otter from Eastern Finland.

Mercury concentrations were monitored in European otter (Lutra lutra), European polecat (Mustela putorius) and European pine marten (Martes martes) collected in Eastern Finland during the period 1972-2008. Otters mainly eat fish, which is an important reason to monitor the bioaccumulation of mercury in this predator. In this species, the highest concentrations were found in fur followed by liver and kidney, and the mercury concentrations increased with increasing age and body weight. Males showed in general higher concentrations than females of otters. The food of European polecat consists of small mammals, frogs, birds and insects from both aquatic and terrestrial food chains. The mercury concentrations were lower than in otters without significant differences related to body weight or sex. In European pine martens, the concentrations were rather evenly distributed except for two specimens with high concentrations. Also, concentrations of some other metals (Al, Cd, Cu, Fe, Mn, Ni, Pb and Zn) were analysed from liver samples of otter. Possible adverse effects of mercury on the Finnish populations of these mustelids are discussed.

Fitting statistical models in bivariate allometry: scaling metabolic rate to body mass in mustelid carnivores.

The ongoing debate about methods for fitting the two-parameter allometric equation y=ax(b) to bivariate data seemed to be resolved recently when three groups of investigators independently reported that statistical models fitted by the traditional allometric method (i.e., by back-transforming a linear model fitted to log-log transformations) typically are superior to models fitted by standard nonlinear regression. However, the narrow focus for the statistical analyses in these investigations compromised the most important of the ensuing conclusions. All the investigations focused on two-parameter power functions and excluded from consideration other simple functions that might better describe pattern in the data; and all relied on Akaike's Information Criterion instead of graphical validation to identify the better statistical model. My re-analysis of data from one of the studies (BMR vs. body mass in mustelid carnivores) revealed (1) that the best descriptor for pattern in the dataset is a straight line and not a two-parameter power function; (2) that a model with additive, normal, heteroscedastic error is superior to one with multiplicative, lognormal, heteroscedastic error; and (3) that Akaike's Information Criterion is not a generally reliable metric for discriminating between models fitted to different distributions. These findings have apparent implications for interpreting the outcomes of all three of the aforementioned studies. Future investigations of allometric variation should adopt a more holistic approach to analysis and not be wedded to the traditional allometric method.

Concentrations of anticoagulant rodenticides in stoats Mustela erminea and weasels Mustela nivalis from Denmark.

Anticoagulant rodenticides are widely used to control rodent populations but they also pose a risk of secondary poisoning in non-target predators. Studies on anticoagulant rodenticide exposure of non-target species have mainly reported on frequency of occurrence. They have rarely analyzed variations in residue concentrations. We examine the occurrence and concentrations of five anticoagulant rodenticides in liver tissue from 61 stoats (Mustela erminea) and 69 weasels (Mustela nivalis) from Denmark. Anticoagulant rodenticides were detected in 97% of stoats and 95% of weasels. 79% of the animals had detectable levels of more than one substance. Difenacoum had the highest prevalence (82% in stoats and 88% in weasels) but bromadiolone was detected in the highest concentrations in both stoat (1.290 µg/g ww) and weasel (1.610 µg/g ww). Anticoagulant rodenticide concentrations were highest during autumn and winter and varied with sampling method. Anticoagulant rodenticide concentrations were higher in stoats and weasels with unknown cause of death than in specimens killed by physical trauma. There was a negative correlation between anticoagulant rodenticide concentrations and body condition. Our results suggest that chemical rodent control in Denmark results in an extensive exposure of non-target species and may adversely affect the fitness of some stoats and weasels.

Effect of energetic constraints on distribution and winter survival of weasel males.

1. The absolute energy needs of small animals are generally lower than those of larger animals. This should drive higher mortality of larger animals, when the environmental conditions deteriorate. However, demonstration of the effect of energy constraints on survivals proved difficult, because the range of body mass within species is generally too small to produce enough variation for studying such an effect. An opportunity for an intraspecific study comes from weasels inhabiting the Bialowieza Forest (north-eastern Poland), which are characterized by a threefold variation in body mass. 2. We assumed that in summer larger weasel males are favoured by sexual selection, because they are more successful when competing for mates. We then tested whether they suffer higher mortality in winter, because they have difficulty finding sufficient food to satisfy their energy needs and/or because the additional foraging time would result in increased exposure to predation. 3. We measured daily energy expenditures (DEE) of overwintering weasel males using the doubly labelled water (DLW) technique. We constructed an energetic model predicting how individuals of different size are able to balance their energy budgets feeding on large and small prey while minimizing time spent hunting, thereby reducing their own exposure to predation. 4. The range of body mass in overwintering weasels predicted by our model corresponded very well with the distribution of prey body mass in three different habitats within our study area. Larger individuals were able to compensate for higher food requirements by using habitats with larger prey species than those available to smaller male weasels. This effectively offset the expected negative association between body mass and winter survival predicted from considerations of energy balance. 5. Our results show how energetic constraints affect body mass and spatial segregation of a species at the intra-specific level not only across large geographical ranges, but also within a relatively small area.

Concentrations of trace elements in tissues of red fox (Vulpes vulpes) and stone marten (Martes foina) from suburban and rural areas in Croatia.

Trace elements concentrations (As, Cd, Cu, Pb and Hg) were determined in the liver, kidney and muscle of 28 red fox (Vulpes vulpes) and 16 stone marten (Martes foina) from suburban and rural habitats from Croatia. Rural and suburban habitats affected Cd and Hg levels in the muscle, liver and kidney of red fox. Significant differences in metal concentrations in the muscle, liver and kidney were detected among species. Suburban stone marten accumulated the highest levels of trace elements (mg/kg w.w.): in muscle 0.019 for Hg; in liver 0.161 for Cd, 36.1 for Cu and 0.349 for Pb; in kidney 1.34 for Cd and 0.318 for Pb. Values observed were higher than those found in suburban red fox and therefore, may represent an important bioindicator for the accumulation of toxic metals in urbanized habitats.

Mortality risk increases with natal dispersal distance in American martens.

The assumption that mortality risk increases with dispersal distance has rarely been tested. We compared patterns of natal dispersal in the American marten (Martes americana) between a large regenerating forest landscape and an uncut landscape that was dominated by more mature forest to test whether mortality risk increased with dispersal distance, and whether variation in mortality risk influenced dispersal distance. Mortality risk increased with dispersal distance in both landscape treatments, but the distance-dependent increase in mortality in the regenerating landscape was twice that in the uncut landscape. Differences in body condition, supported by other data on foraging efficiency, suggested that juveniles from the regenerating landscape were less able to cope with the energetic demands of dispersal compared with juveniles from older forests. Juveniles travelled shorter distances in the regenerating versus uncut landscape. These results implied that dispersal was costly in terms of juvenile survival and that mean dispersal distance was shaped, in part, by mortality risk.

Unusual odd-chain and trans-octadecenoic fatty acids in tissues of feral European beaver (Castorfiber), Eurasian badger (Melesmeles) and raccoon dog (Nyctereutesprocyonoides).

The fatty acid (FA) composition of depot adipose tissues in the raccoon dog (Nyctereutesprocyonoides) and the European beaver (Castorfiber) differs from that reported for the lipids of other monogastric animals, especially with regard to the presence of trans-octadecenoic acids. The concentrations of pentadecanoic acid 15:0 (PA) and heptadecanoic acid 17:0 (HA) in the lipids of the tested animals ranged from 0.23 to 0.79% and from 0.33 to 2.35% of total FAs, respectively. The total content of their monounsaturated cis isomers varied from 0.12 to 2.75% for pentadecanoic acid (c-PA) and from 0.38 to 2.45% for heptadecanoic acid (c-HA). It is interesting that the tissues of European beavers and raccoon dogs contained also trans isomers of octadecenoic acid C18:1 (t-OA) including vaccenic acid C18:1,11t (VA), typical of ruminants. The presence of FAs with an uneven number of carbon atoms and trans-octadecenoic acids in depot adipose tissue is indicative of the process of hydrogenation of polyunsaturated fatty acids (linoleic acid and alpha-linolenic acid) in the digestive tract. The tissues of badgers also contained t-OA (from below 0.05% in the liver to 0.44% in the kidneys), but no VA was found.

Fatty acid profiles and relative mobilization during fasting in adipose tissue depots of the American marten (Martes americana).

The American marten (Martes americana) is a boreal forest marten with low body adiposity but high metabolic rate. The study describes the FA composition in white adipose tissue depots of the species and the influence of food deprivation on them. American marten (n = 8) were fasted for 2 d with 7 control animals. Fasting resulted in a 13.4% weight loss, while the relative fat mass was >25% lower in the fasted animals. The FA composition of the fat depots of the trunk was quite similar to other previously studied mustelids with 14:0, 16:0, 18:0, 16:1 n-7, 18:1 n-9, and 18:2n-6 as the most abundant FA. In the extremities, there were higher proportions of monounsaturated FA (MUFA) and PUFA. Food deprivation decreased the proportions of 16:0 and 16:1 n-7, while the proportion of long-chain MUFA increased in the trunk. The mobilization of FA was selective, as 16:1 n-7, 18:1 n-9, and particular n-3 PUFA were preferentially mobilized. Relative mobilization correlated negatively with the carbon chain length in saturated FA (SFA) and n-9 MUFA. The delta9-desaturation of SFA enhanced the mobilization of the corresponding MUFA, but the positional isomerism of the first double bond did not correlate consistently with relative mobilization in MUFA or PUFA. In the marten, the FA composition of the extremities was highly resistant to fasting, and the tail tip and the paws contained more long-chain PUFA to prevent the solidification of lipids and to maintain cell membrane fluidity during cooling.

Long-lasting systemic bait markers for Eurasian badgers.

This study was carried out to assess whether Rhodamine B, ethyl-iophenoxic acid (EtIPA), and propyl-iophenoxic acid (PrIPA) can be used as long-lasting systemic bait markers for free-living badgers (Meles meles). Between June and November 2003, these chemicals were incorporated into bait distributed around badger setts. Serum, hair, and whiskers from individually marked badgers were collected in the following 4 to 24 wk. Rhodamine B was detectable as fluorescent bands up to 24 wk after ingestion of the bait. Individual badgers were found positive for EtIPA and PrIPA up to 20 wk and 18 wk after exposure, respectively. This study indicates that Rhodamine B, PrIPA, and EtIPA could be used as long-lasting markers for badgers.